The invention relates generally to transforming growth factor alpha ( ) and more specifically to methods of using TGF-xcex1 for stimulating hematopoiesis, for suppressing immune function associated with autoimmune diseases, for suppressing inflammatory responses mediated by excessive histamine release and by expression of TNF-receptors and associated pro-inflammatory cytokines, and for treating cachexia or for treating or preventing mucositis and gastrointestinal-associated disorders.
There are several disease treatments that could significantly benefit by having cells regenerate after injury or lesion formation, particularly in the CNS, in the immune system and in the gastrointestinal tract. In some instances, a particular treatment for a disease often detrimentally affects the subject being treated. For example, administration of chemotherapeutic agents to subjects results in destruction of healthy cells, for example, cells of the gastrointestinal tract. A number treatment-related disorders are related to the choice of chemotherapeutic agent. Such agents include carmustine (BCNU), chlorambucil (LEUKERAN), cisplatin (PLATINOL), Cytarabine, doxorubicin (ADRIAMYCIN), fluorouracil (5-FU), methoxetrate (MEXATE), taxol, CPT111, etoposide, and plicamycin (MITHRACIN) which are known for their direct stomatotoxic potential (Sonis, 1993, xe2x80x9cOral Complications in Cancer Therapy,xe2x80x9d In: Principles and Practice of Oncology, pp. 2385-2394, DeVitta et al., Eds., J. B. Lippincott, Philadelphia) and hence incidence of mucositis.
Oral mucositis can be initiated by the cytotoxic effects of chemotherapy and/or radiotherapy on the rapidly dividing epithelial cells of the oropharyngeal mucosa, and is exacerbated by infection with both endogenous oral flora and opportunistic bacterial and fungal pathogens. Complications related to oral mucositis vary in the different patient populations affected, but typically include pain, poor oral intake with consequent dehydration and weight loss, and systemic infection with organisms originating in the oral cavity. The pain associated with oral mucositis may be severe requiring narcotic analgesics, and the difficulty in eating can result in patients receiving total parenteral nutrition.
Current therapies have been directed at decreasing oral flora and the extent of infection of oral ulcerations. Systemic treatment with G- and GM-CSF has been shown to result in a decreased incidence of oral mucositis, presumably by allowing for more rapid neutrophil recovery and thus an improved ability to combat infection, although it has been postulated that the CSFs may have a more direct effect on the oral mucosa (Chi et al., 1995, J. Clin. Oncol. 13:2620-2628). In one study, GM-CSF was reported to exacerbate mucositis. (Cartee et al., 1994, Cytokine 7:471-477). Benzydamine hydrochloride, a nonsteroidal drug with analgesic and antimicrobial properties, has been studied both in patients undergoing radiation therapy and in patients receiving intra-arterial chemotherapy.
In addition, diseases associated with epithelial cell depletion in the gastrointestinal tract often increase the risk of related disorders. Such related disorders include infection by opportunistic pathogens as well as weight loss associated with the loss in nutrient uptake in the gastrointestinal tract.
The present invention is based on the seminal discovery that TGF-xcex1 and functionally related polypeptides, TGF-xcex1 mimetics, functional TGF-xcex1 peptides, and polynucleotides encoding such polypeptides and peptide fragments are effective for treating or preventing weight-loss in subjects having disorders or diseases associated with weight-loss (e.g., cachexia). In addition, the polypeptides of the invention have mitogenic and barrier function (e.g., protective) effects on stem cells and their differentiated progeny from a variety of tissues including the gastrointestinal system, the nervous system and the hematopoietic system.
In a first embodiment, the invention provides a method of treating a subject having or at risk of having cachexia comprising administering to the subject a transforming growth factor-alpha (TGF-xcex1) polypeptide in an amount effective to prevent or reduce weight-loss. In one aspect, the invention provides a method of increasing the body weight of a subject comprising administering to the subject, prior to, simultaneously with, or substantially following chemotherapy, a transforming growth factor-alpha (TGF-xcex1) polypeptide in an amount effective to increase the weight of the subject. In one aspect the subject has AIDS related complex (ARC) or AIDS and cachexia associated with such diseases.
In another embodiment, the invention provides a method for treating or preventing mucositis of the gastrointestinal tract in a subject, comprising administering a TGFxcex1 or related polypeptide in an amount effective to treat or prevent mucositis in the subject. For example, a subject undergoing chemotherapy can be treated by the method of the invention.
The invention also provides a polypeptide comprising a peptide having a sequence NH2-X1a-Cys-His-Ser-X1b-X2-X1a-X1b-X1a-X3-Cys-COOH (SEQ ID NO:4) wherein X1a and X1b are independently Val, Gly or Ala; X2 is Tyr or Phe; X3 is Arg or Lys; and the two Cys moieties are linked via a disulfide bond to form an at least 11-amino acid functional peptide having TGF-xcex1 activity.
In another embodiment, the invention provides a polypeptide comprising a peptide having a sequence NH2-X1a-Cys-His-Ser-X1b-X2-X1a-X1b-X1a-X3-Cys-COOH (SEQ ID NO:4) wherein X1a and X1b are independently Val, Gly or Ala; X2 is Tyr or Phe; X3 is Arg or Lys; and the two Cys moieties are linked via a disulfide bond to form an at least 11-amino acid functional peptide having TGF-xcex1 activity and wherein at least one or more of the following amino acids are linked to the C-terminal Cys moiety of SEQ ID NO:4: -X4-His-X1c-X4-X5-X6-X1c (SEQ ID NO:5) wherein X4 is Glu or Asp; X5 is Leu or Ile; and X6 is Asp or Glu.
In yet another embodiment, the invention provides a pharmaceutical composition comprising a polypeptide having a sequence NH2-X1a-Cys-His-Ser-X1b-X2-X1a-X1b-X1a-X3-Cys COOH (SEQ ID NO:4) wherein X1a and X1b are independently Val, Gly or Ala; X2 is Tyr or Phe; X3 is Arg or Lys; and the two Cys moieties are linked via a disulfide bond to form an at least 11-amino acid functional peptide having TGF-xcex1 activity, and a pharmaceutically acceptable carrier. In addition, at least one or more of the following amino acids are linked to the C-terminal Cys moiety of SEQ ID NO:4: -X4-His-X1c-X4-X5-X6-X1c (SEQ ID NO:5) wherein X4 is Glu or Asp; X5 is Leu or Ile; and X6 is Asp or Glu.
The invention also provides a compound that acts as a TGF-xcex1 mimetic, comprising a compound of formula: loop peptide N-terminus-linker-cyclic C4H8N2-linker-loop peptide N-terminus wherein the linker moiety is designed to link the N-terminus of the loop peptide to a nitrogen atom of the ring C4H8N2 and wherein the loop peptide has a sequence NH2-X1a-Cys-His-Ser-X1b-X2-X1a-X1b-X1a-X3-Cys COOH (SEQ ID NO:4) wherein X1a and X1b are independently Val, Gly or Ala; X2 is Tyr or Phe; X3 is Arg or Lys; and the two Cys moieties are linked via a disulfide bond to form an at least 11-amino acid functional peptide having TGF-xcex1 activity.